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An IL-9 fate reporter demonstrates the induction of an innate IL-9 response in lung inflammation.

Wilhelm C, Hirota K, Stieglitz B, Van Snick J, Tolaini M, Lahl K, Sparwasser T, Helmby H, Stockinger B - Nat. Immunol. (2011)

Bottom Line: We found that during papain-induced lung inflammation, IL-9 production was largely restricted to innate lymphoid cells (ILCs).IL-9 production by ILCs depended on IL-2 from adaptive immune cells and was rapidly lost in favor of other cytokines, such as IL-13 and IL-5.Blockade of IL-9 production via neutralizing antibodies resulted in much lower expression of IL-13 and IL-5, which suggested that ILCs provide the missing link between the well-established functions of IL-9 in the regulation of type 2 helper T cell cytokines and responses.

View Article: PubMed Central - PubMed

Affiliation: Division of Molecular Immunology, Medical Research Council National Institute for Medical Research, Mill Hill, UK.

ABSTRACT
Interleukin 9 (IL-9) is a cytokine linked to lung inflammation, but its cellular origin and function remain unclear. Here we describe a reporter mouse strain designed to map the fate of cells that have activated IL-9. We found that during papain-induced lung inflammation, IL-9 production was largely restricted to innate lymphoid cells (ILCs). IL-9 production by ILCs depended on IL-2 from adaptive immune cells and was rapidly lost in favor of other cytokines, such as IL-13 and IL-5. Blockade of IL-9 production via neutralizing antibodies resulted in much lower expression of IL-13 and IL-5, which suggested that ILCs provide the missing link between the well-established functions of IL-9 in the regulation of type 2 helper T cell cytokines and responses.

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ILC are the major source of IL-9a) Flow cytometry of lung cells isolated from papain challenged B6 mice 12 h after papain re-challenge, restimulated with PdBU and ionomycin for 2.5 h and stained for lineage markers and intracellular IL-9 and IL-13. b) Intracellular IL-9 and IL-13 expression in gated CD4+ cells or ILC. c) Absolute number of ILC and CD4+ cells and d) IL9+IL13+ ILC or CD4+ cells in the lungs of papain challenged mice 12 h after re-challenge. e) Flow cytometry of lung cells isolated from Rag2−/−IL2rg−/− mice (no transfer) or Rag2−/−IL2rg−/− mice transferred with CD4+ cells or ILC and challenged with papain and IL-2 (see scheme Supplementary Fig.9). Lung cells were stained for CD45 and Thy1.2 (left panel), restimulated with PdBU and ionomycin for 2.5 h, stained for intracellular IL-9 and IL-13 and gated on CD45+ Thy1.2+ cells (middle panel). Right panel: Absolute number of IL-9+IL-13+ cells (p= 0.0238). f) Cytokine concentration in the lung homogenate of papain and IL-2 challenged Rag2−/−IL2rg−/− mice (white bars) or papain and IL-2 challenged Rag2−/−IL2rg−/− mice transferred with CD4+ cells (grey bars) or ILC (black bars) 15 h after papain challenge. Numbers in gates or quadrants (a, b, e) indicate percent cells in each. Data represents at least two independent experiments with 3 mice in each experimental group (mean ±SEM in c, d, e and f).
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Figure 6: ILC are the major source of IL-9a) Flow cytometry of lung cells isolated from papain challenged B6 mice 12 h after papain re-challenge, restimulated with PdBU and ionomycin for 2.5 h and stained for lineage markers and intracellular IL-9 and IL-13. b) Intracellular IL-9 and IL-13 expression in gated CD4+ cells or ILC. c) Absolute number of ILC and CD4+ cells and d) IL9+IL13+ ILC or CD4+ cells in the lungs of papain challenged mice 12 h after re-challenge. e) Flow cytometry of lung cells isolated from Rag2−/−IL2rg−/− mice (no transfer) or Rag2−/−IL2rg−/− mice transferred with CD4+ cells or ILC and challenged with papain and IL-2 (see scheme Supplementary Fig.9). Lung cells were stained for CD45 and Thy1.2 (left panel), restimulated with PdBU and ionomycin for 2.5 h, stained for intracellular IL-9 and IL-13 and gated on CD45+ Thy1.2+ cells (middle panel). Right panel: Absolute number of IL-9+IL-13+ cells (p= 0.0238). f) Cytokine concentration in the lung homogenate of papain and IL-2 challenged Rag2−/−IL2rg−/− mice (white bars) or papain and IL-2 challenged Rag2−/−IL2rg−/− mice transferred with CD4+ cells (grey bars) or ILC (black bars) 15 h after papain challenge. Numbers in gates or quadrants (a, b, e) indicate percent cells in each. Data represents at least two independent experiments with 3 mice in each experimental group (mean ±SEM in c, d, e and f).

Mentions: To investigate whether ILC are the critical producers of IL-9 during papain-induced lung inflammation we performed intracellular cytokine staining for IL-9 and IL-13 in ILC and CD4+ T cells. The majority of intracellular IL-9 was expressed in ILC, although we detected a minor contribution from CD4+ T cells (Fig. 6a, b). While similar numbers of ILC and CD4+ T cells are recovered from the lungs of papain-challenged mice (Fig. 6c), IL-9 and IL-13 expression stems almost exclusively from ILC (Fig. 6d).


An IL-9 fate reporter demonstrates the induction of an innate IL-9 response in lung inflammation.

Wilhelm C, Hirota K, Stieglitz B, Van Snick J, Tolaini M, Lahl K, Sparwasser T, Helmby H, Stockinger B - Nat. Immunol. (2011)

ILC are the major source of IL-9a) Flow cytometry of lung cells isolated from papain challenged B6 mice 12 h after papain re-challenge, restimulated with PdBU and ionomycin for 2.5 h and stained for lineage markers and intracellular IL-9 and IL-13. b) Intracellular IL-9 and IL-13 expression in gated CD4+ cells or ILC. c) Absolute number of ILC and CD4+ cells and d) IL9+IL13+ ILC or CD4+ cells in the lungs of papain challenged mice 12 h after re-challenge. e) Flow cytometry of lung cells isolated from Rag2−/−IL2rg−/− mice (no transfer) or Rag2−/−IL2rg−/− mice transferred with CD4+ cells or ILC and challenged with papain and IL-2 (see scheme Supplementary Fig.9). Lung cells were stained for CD45 and Thy1.2 (left panel), restimulated with PdBU and ionomycin for 2.5 h, stained for intracellular IL-9 and IL-13 and gated on CD45+ Thy1.2+ cells (middle panel). Right panel: Absolute number of IL-9+IL-13+ cells (p= 0.0238). f) Cytokine concentration in the lung homogenate of papain and IL-2 challenged Rag2−/−IL2rg−/− mice (white bars) or papain and IL-2 challenged Rag2−/−IL2rg−/− mice transferred with CD4+ cells (grey bars) or ILC (black bars) 15 h after papain challenge. Numbers in gates or quadrants (a, b, e) indicate percent cells in each. Data represents at least two independent experiments with 3 mice in each experimental group (mean ±SEM in c, d, e and f).
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Figure 6: ILC are the major source of IL-9a) Flow cytometry of lung cells isolated from papain challenged B6 mice 12 h after papain re-challenge, restimulated with PdBU and ionomycin for 2.5 h and stained for lineage markers and intracellular IL-9 and IL-13. b) Intracellular IL-9 and IL-13 expression in gated CD4+ cells or ILC. c) Absolute number of ILC and CD4+ cells and d) IL9+IL13+ ILC or CD4+ cells in the lungs of papain challenged mice 12 h after re-challenge. e) Flow cytometry of lung cells isolated from Rag2−/−IL2rg−/− mice (no transfer) or Rag2−/−IL2rg−/− mice transferred with CD4+ cells or ILC and challenged with papain and IL-2 (see scheme Supplementary Fig.9). Lung cells were stained for CD45 and Thy1.2 (left panel), restimulated with PdBU and ionomycin for 2.5 h, stained for intracellular IL-9 and IL-13 and gated on CD45+ Thy1.2+ cells (middle panel). Right panel: Absolute number of IL-9+IL-13+ cells (p= 0.0238). f) Cytokine concentration in the lung homogenate of papain and IL-2 challenged Rag2−/−IL2rg−/− mice (white bars) or papain and IL-2 challenged Rag2−/−IL2rg−/− mice transferred with CD4+ cells (grey bars) or ILC (black bars) 15 h after papain challenge. Numbers in gates or quadrants (a, b, e) indicate percent cells in each. Data represents at least two independent experiments with 3 mice in each experimental group (mean ±SEM in c, d, e and f).
Mentions: To investigate whether ILC are the critical producers of IL-9 during papain-induced lung inflammation we performed intracellular cytokine staining for IL-9 and IL-13 in ILC and CD4+ T cells. The majority of intracellular IL-9 was expressed in ILC, although we detected a minor contribution from CD4+ T cells (Fig. 6a, b). While similar numbers of ILC and CD4+ T cells are recovered from the lungs of papain-challenged mice (Fig. 6c), IL-9 and IL-13 expression stems almost exclusively from ILC (Fig. 6d).

Bottom Line: We found that during papain-induced lung inflammation, IL-9 production was largely restricted to innate lymphoid cells (ILCs).IL-9 production by ILCs depended on IL-2 from adaptive immune cells and was rapidly lost in favor of other cytokines, such as IL-13 and IL-5.Blockade of IL-9 production via neutralizing antibodies resulted in much lower expression of IL-13 and IL-5, which suggested that ILCs provide the missing link between the well-established functions of IL-9 in the regulation of type 2 helper T cell cytokines and responses.

View Article: PubMed Central - PubMed

Affiliation: Division of Molecular Immunology, Medical Research Council National Institute for Medical Research, Mill Hill, UK.

ABSTRACT
Interleukin 9 (IL-9) is a cytokine linked to lung inflammation, but its cellular origin and function remain unclear. Here we describe a reporter mouse strain designed to map the fate of cells that have activated IL-9. We found that during papain-induced lung inflammation, IL-9 production was largely restricted to innate lymphoid cells (ILCs). IL-9 production by ILCs depended on IL-2 from adaptive immune cells and was rapidly lost in favor of other cytokines, such as IL-13 and IL-5. Blockade of IL-9 production via neutralizing antibodies resulted in much lower expression of IL-13 and IL-5, which suggested that ILCs provide the missing link between the well-established functions of IL-9 in the regulation of type 2 helper T cell cytokines and responses.

Show MeSH
Related in: MedlinePlus